Ishii Yuki, Orr Jessica C, El Mdawar Marie-Belle, de Pilger Denise R Bairros, Pearce David R, Lazarus Kyren A, Graham Rebecca E, Nikolić Marko Z, Ketteler Robin, Carragher Neil O, Janes Sam M, Hynds Robert E
Lungs for Living Research Centre, UCL Respiratory, University College London, London WC1E 6JF, UK.
Epithelial Cell Biology in ENT Research Group, Developmental Biology and Cancer Department, UCL Great Ormond Street Institute of Child Health, University College London, London WC1N 1DZ, UK.
J Cell Sci. 2025 Apr 1;138(7). doi: 10.1242/jcs.263487. Epub 2025 Apr 14.
Regeneration of the airway epithelium restores barrier function and mucociliary clearance following lung injury and infection. The mechanisms regulating the proliferation and differentiation of tissue-resident airway basal stem cells remain incompletely understood. To identify compounds that promote human airway basal cell proliferation, we performed phenotype-based compound screening of 1429 compounds (from the ENZO and Prestwick Chemical libraries) in 384-well format using primary cells transduced with lentiviral luciferase. A total of 17 pro-proliferative compounds were validated in independent donor cell cultures, including the antiretroviral therapy agent abacavir and several Wnt signalling pathway-activating compounds. The effects of compounds on proliferation were further explored in colony formation and 3D organoid assays. Structurally and functionally related compounds that more potently induced Wnt pathway activation were investigated. One such compound, 1-azakenpaullone, induced Wnt target gene activation and basal cell proliferation in mice. Our results demonstrate the pro-proliferative effect of small-molecule Wnt pathway activators on airway basal cells. These findings contribute to the rationale to develop novel approaches to modulate Wnt signalling during airway epithelial repair.
气道上皮的再生可在肺损伤和感染后恢复屏障功能和黏液纤毛清除功能。调节组织驻留气道基底干细胞增殖和分化的机制仍未完全了解。为了鉴定促进人气道基底细胞增殖的化合物，我们使用慢病毒荧光素酶转导的原代细胞，以384孔板形式对1429种化合物（来自ENZO和Prestwick Chemical文库）进行了基于表型的化合物筛选。在独立供体细胞培养物中验证了总共17种促增殖化合物，包括抗逆转录病毒治疗药物阿巴卡韦和几种Wnt信号通路激活化合物。在集落形成和3D类器官测定中进一步探索了化合物对增殖的影响。研究了更有效诱导Wnt途径激活的结构和功能相关化合物。一种这样的化合物，1-氮杂肯帕罗酮，在小鼠中诱导Wnt靶基因激活和基底细胞增殖。我们的结果证明了小分子Wnt途径激活剂对气道基底细胞的促增殖作用。这些发现为开发在气道上皮修复过程中调节Wnt信号传导的新方法提供了理论依据。
